Abstract There has been the misconception that rabies does not pose a threat to human health anymore due to the highly effective rabies virus vaccine available for prevention and post-exposure treatment. However, it has been overlooked that the current rabies virus vaccine only protects against lyssaviruses of the phylogroup I and not against other, emerging lyssaviruses. Moreover, protection against some members of phylogroup I is less than optimal and requires high antibody titers. This proposal seeks to eventually shift clinical practice paradigms by offering a vaccine with more comprehensive protection against rabies from divergent lyssaviruses. Toward this approach, we will construct novel rabies-virus-based vectors expressing glycoproteins of other lyssaviruses, immunize mice and then characterize the humoral immune response on an antigen microarray of overlapping peptides spanning each of the glycoproteins. The results of these studies will guide us to construct novel G proteins, which should induce a broader immune response against lyssavirus and therefore pave the way toward a novel pan-lyssavirus vaccine.